STUDIES  ON  WATER-DRINKING 


VI.  THE  ACTIVITY  OF  THE  PANCREATIC  FUNCTION  UNDER 
THE  INFLUENCE  OF  COPIOUS  AND  MODERATE 
WATER-DRINKING  WITH  MEALS 


P.  B.  H A W K,  Ph.D. 


URBANA,  ILL. 


REMOTE  s t 


6/2-. 3 
H 3 1 a- 


Crp  p/  2~ 
(^Js^Qaas  ~ * - 


STUDIES  ON  WATER-DRINKING 


VI.  the  activity  of  the  pancreatic  function  under  the  influence 

OF  COPIOUS  AND  MODERATE  WATER-DRINKING  WITH  MEALS  *  *  1 


P.  B.  HAWK,  Ph.D. 

TJBBANA,  ILL. 


Numerous  tests  are  in  vogue  clinically  for  the  determination  of  the 
functional  activity  of  the  pancreas.  The  best  known  of  these  are  the 
glutoid  test  of  Sahli,2  the  nuclei  test  of  Schmidt,3  and  the  Cammidge4 
reaction,  the  last  mentioned  test  having  been  rather  more  freely  criticized 
than  either  of  the  others.  Comparatively  recently,  clinicians  have  come 
to  a fuller  realization  of  the  aid  which  a knowledge  of  the  pancreatic 
enzyme  concentration  of  the  feces  may  give  them  in  their  diagnosis  of 
pancreatic  insufficiency.  The  accurate  determination  of  the  enzyme 
content  of  the  feces  under  normal  or  pathological  conditions  is  still,  how- 
ever, a line  of  medical  research  which  has  not  been  very  generally  investi- 
gated. Some  of  the  investigators  who  have  recently  contributed  to  our 
knowledge  in  this  direction  either  by  suggestions  as  to  method  or  by  the 
collection  of  clinical  data  are  Muller,5  Wohlgemuth,6  Wynhausen,7  Boldy- 
reff,8  Abderhalden  and  Schittenhelm,9  Schlecht,10  Goldschmidt,11  Vol- 
hard,12  Ury,13  Frank  and  Schittenhelm.14  One  of  the  first  enzymes  to  be 

*From  the  Laboratory  of  Physiological  Chemistry  of  the  University  of 
Illinois. 

1.  For  previous  papers  in  this  series  of  studies  see,  I.  Hawk:  Univ.  Penn. 
Med.  Bull.,  1905,  xviii,  7;  II.  Fowler  and  Hawk:  Jour.  Exper.  Med.,  1910,  xii, 
388;  III.  Rulon  and  Hawk:  Jour.  Am.  Chem.  Soc.,  1910,  xxxii,  1686;  IV.  Rulon 
and  Hawk:  The  Archives  Int.  Med.,  1911,  vii,  536;  V.  Hattrem  and  Hawk: 
Ti-ie  Archives  Int.  Med.  1911,  vii,  610. 

2.  Sahli:  Deutsch.  Arch.  f.  klin.  Med.,  1898,  p.  61. 

3.  Schmidt:  Deutsch.  med.  Wchnschr.,  1899,  No.  45. 

4.  Cammidge:  Proc.  Roy.  Soc.,  London  (B),  1909,  lxxxi,  372;  also  Proc. 
Roy.  Soc.  Med.,  iii,  164. 

5.  Muller:  Arch.  f.  klin.  Med.,  1908. 

6.  Wohlgemuth:  Berl.  klin.  Wchnschr.,  1910,  xlvii,  92. 

7.  Wynhausen:  Berl.  klin.  Wchnschr.,  1909,  xlvi,  1406. 

8.  Boldyreff:  Arch.  f.  d.  ges.  Physiol.  (Pfliigers),  1905,  cxxi,  13. 

9.  Abderhalden  and  Schittenhelm:  Ztschr.  f.  physiol.  Chem.,  1909,  lix,  230. 

10.  Schlecht:  Miinchen.  med.  Wchnschr.,  1908,  lv,  725. 

11.  Goldschmidt:  Deutsch.  med.  Wchnschr.,  1909,  No.  12. 

12.  Volhard:  Miinchen.  med.  Wchnschr.,  1908,  liv,  403. 

13.  Ury:  Biochem.  Ztschr.,  1909,  xxiii,  153. 

14.  Frank  and  Schittenhelm:  Ztschr.  f.  exper.  Path.  u.  Therap.,  1910,  viii, 
237. 


T3  36722 


4 


detected  in  the  feces  was  pancreatic  amylase.  This  enzyme,  as  early  as 
1875,  was  shown  by  Wegscheider15  to  be  present  in  the  feces  of  the 
nursling.  This  finding  was  later  verified  by  von  Jaksch,16  Moro17  and 
Allaria.18  According  to  this  last-mentioned  investigator  the  manner  of 
feeding  the  infant,  i.  e.,  naturally  or  artificially,  had  no  influence  on  the 
amylase  concentration  of  the  feces.  Strasburger19  found  this  same 
condition  to  hold  for  the  stools  of  adults. 

DESCRIPTION'  OF  EXPERIMENTS 

Three  experiments  were  made,  in  each  of  which  normal  men  (E  and 
W)  served  as  subjects,  the  same  man  serving  as  subject  in  Experiments 
2 and  3.  Each  experiment  was  divided  into  three  periods,  a fore  period 
during  which  the  subject  was  brought  into  nitrogen  equilibrium  through 
the  ingestion  of  a diet  uniform  as  to  quality  and  quantity  from  day  to 
day,  a water  period  during  which  the  ration  was  supplemented  by  the 
drinking  of  additional  water  at  meal-time,  and  finally  an  after  period  in 
which  the  fore  period  ration  of  low  water  content  was  again  fed.  The 
constituents  of  the  diet  apart  from  the  water  were  graham  crackers,  butter, 
milk  and -peanut  butter.  The  diet  of  Subject  W in  Experiment  1 con- 
tained 85.8  gm.  of  protein  per  day  whereas  that  of  Subject  E in  Experi- 
ments 2 and  3 contained  92.3  gm.  of  protein  per  day.  During  the  fore 
and  after  periods  of  each  experiment  100  c.c.  of  water,  in  addition  to  the 
water  content  of  the  milk,  was  taken  by  each  subject  at  each  meal.  In  the 
water  periods  of  Experiments  1 and  2,  500  c.c.  of  water  above  that  previ- 
ously ingested  were  taken  with  each  meal,  whereas  in  Experiment  3 this 
volume  was  increased  to  1%  liters. 

Each  individual  stool  was  examined  in  the  fresh  condition.  The  feces 
of  one  period  were  separated  from  those  of  the  next  by  means  of  charcoal, 
the  material  being  ingested  in  gelatin  capsules  at  the  beginning  of  the 
first  meal  of  each  new  period. 

The  method  used  in  the  quantitative  determination  of  the  activity  of 
the  pancreatic  function  was  that  suggested  by  Wohlgemuth.6  This  author 
considers  that  the  extent  to  which  pancreatic  amylase  appears  in  the  feces 
may  be  taken  as  an  index  of  the  activity  of  the  pancreatic  function.  He 
therefore  suggests  a procedure  for  the  quantitative  determination  of  the 
amylolytic  power  of  sodium  chlorid  extracts  of  the  fresh  feces.  His 
procedure  has  been  fully  discussed  by  us  in  another  connection  and  a 
modification  of  the  Wohlgemuth  method  proposed.20  In  the  experiments 

15.  Wegscheider:  Inaug.  Diss.,  Strassburg,  1875. 

16.  Von  Jaksch:  Ztschr.  f.  physiol.  Chem.,  1888,  xii,  116. 

17.  Moro:  Jahrb.  f.  Kinderh.,  1898,  xlvii,  342. 

18.  Allaria:  Progresso  Med.,  1905. 

19.  Strasburger:  Arch.  klin.  Med.,  1900,  lxvii,  238  and  531. 

20.  Hawk:  The  Archives  Int.  Med.;  to  be  publisned. 


5 


embraced  in  the  present  paper,  however,  the  procedure  used  by  ns  was 
essentially  that  of  Wohlgemuth  except  that  the  final  amylolytic  values  of 
the  stools  were  calculated  in  terms  of  dry  matter  instead  of  on  the  basis 
of  the  moist  sediment  secured  by  centrifugation  as  Wohlgemuth  suggests. 

In  order  to  demonstrate  conclusively  the  origin  of  the  fecal  amylase 
Wohlgemuth6  has  made  certain  interesting  tests.  In  these  tests  it  was 
demonstrated  that  the  ligation  of  the  pancreatic  ducts  of  dogs  caused  the 
feces  as  passed  to  be  amylase-free  or  to  contain  at  the  most  a minimal 
quantity  of  the  enzyme.  Wynhausen21  has  likewise  found  a much- 
decreased  amylase  output  in  cases  with  partial  closure  of  the  pancreatic 
duct. 

Table  1. — Fecal  Amylase,  Experiment  1,  Subject  W,  Moderate  Water-Drinking 


FORE  PERIOD SIX  DAYS 


Feces  — 

Amylolytic 

Value — 1 °fo  Starch 

Stool 

Fresh 

Dry 

Total 

Per  Gram  Dry 

No. 

(grams) 

(grams) 

(liters) 

Matter  (c.c.) 

1 

62.8 

16.1 

11.4 

709 

2 

98.8 

26.4 

10.8 

409 

3 

109.8 

30.2 

18.1 

600 

4 

185.8 

44.8 

26.3 

587 

5 

41.8 

13.1 

6.7 

513 

6 

104.9 

25.5 

20.9 

819 

7 

24.0 

7.9 

3.3 

416 

Average  per  day. . . 

. . 104.6 

27.3 

16.3 

597 

WATER  PERIOD TEN  DAYS 

(1,500  c.c.  HoO  daily) 

8 

31.6 

7.6 

7.6 

1,000 

9 

147.5 

40.6 

34.9 

860 

10 

75.4 

19.8 

17.1 

865 

11 

144.8 

39.8 

27.3 

688 

12 

63.9 

17.2 

14.6 

848 

13 

115.5 

31.3 

22.2 

709 

14 

26.0 

7.2 

4.0 

548 

15 

169.0 

44.8 

32.5 

725 

16 

127.0 

30.9 

44.7 

1,447 

17 

152.7 

35.5 

110.3 

3,106 

Average  per  dav. . . 

. . 105.3 

27.5 

31.5 

1,145 

AFTER  PERIOD 

18 

60.0 

14.8 

14.6 

983 

19 

119.6 

28.0 

29.4 

1,052 

20 

51.1 

14.1 

20.8 

1,477 

21 

142.7 

35.8 

41.1 

1,147 

22 

81.4 

22.8 

15.0 

657 

23 

53.4 

16.4 

19.1 

1,162 

Average  per  day. . . 

. . 101.6 

26.3 

28.0 

1,065 

DISCUSSION  OF  RESULTS 

1.  Moderate  W at er -Drinking . — Data  from  the  first  experiment  may  be 
found  in  Table  1.  The  subject  of  this  experiment  was  W and  the  purpose 
was  the  study  of  the  influence  of  moderate  water-drinking  with  meals 
on  the  activity  of  the  pancreatic  function.  After  a fore  period  of  six 
days  on  a uniform  diet,  as  already  mentioned,  the  subject  was  found  to  be 
in  nitrogen  equilibrium  as  was  shown  by  a nitrogen  balance  of  0.01  + 
gm.  The  average  daily  output  of  fresh  feces  was  104.6  gm.,  representing 
a dry  matter  content  of  27.3  gm.  per  day  during  this  period.22  The  total 


21.  Wynhausen:  Berl.  klin.  Wchnschr.,  1909,  No.  30. 

22.  The  data  for  fresh  and  dry  feces  were  obtained  by  Dr.  H.  A.  Mattill  in 
connection  with  another  research  which  has  already  been  reported.  See  Mattill 
and  Hawk:  Proc.  Soc.  Biol.  Chem.,  1911. 


amylolytic  value  of  the  stools  per  day,  expressed  in  liters  of  1 per  cent, 
starch  solution  which  this  feces  would  hydrolyze  under  the  conditions  of 
the  Wohlgemuth  method,  was  16.3  liters.  Placed  on  the  basis  of  “1  gm. 
of  dr}^  feces”  we  see  by  an  examination  of  the  data  that  the  average  daily 
amylolytic  value  for  this  period  was  represented  by  597  c.c.  of  1 per  cent, 
starch  solution. 

Under  the  influence  of  the  500  c.c.  of  water  ingested  at  each  meal  for 
a period  of  ten  days  there  was  a marked  increase  in  the  amylolytic  power 
of  the  feces.  For  example,  the  data  from  the  first  stool  of  this  period 
indicate  that  the  amylolytic  value  was  1,000  c.c.  of  1 per  cent,  starch 
solution  per  gram  of  dry  feces.  This  initial  value  of  the  period  was  far 
above  that  for  any  individual  stool  of  the  fore  period  and  nearly  double 
that  of  the  daily  average  amylolytic  value  (597  c.c.)  for  the  entire  fore 
period.  If  the  data  from  the  analysis  of  the  stools  which  were  subse- 
quently dropped  during  this  water  period  are  examined  it  will  be  seen 
that  the  amylolytic  values  for  these  stools,  with  a single  exception 
(No.  14),  were  above  the  average  value  for  the  fore  period.  It  will 
furthermore  be  observed  that  the  average  amylolytic  value  for  the  water 
period,  i.  e.,  1,145,  was  approximately  twice  as  great  as  the  average  amylo- 
lytic value  for  the  fore  period.  In  other  words  the  drinking  of  1,500  c.c. 
of  water  per  day,  at  meal  time,  through  a period  of  ten  days,  was  instru- 
mental in  causing  a virtual  doubling  of  the  anxiolytic  activity  of  the  fecal 
matter  excreted  during  that  period  as  compared  with  the  fecal  output  of 
the  fore  period  during  which  minimal  amounts  of  water  were  daily 
ingested. 

The  significance  of  this  finding  of  an  increased  amylolytic  power  is 
further  accentuated  when  the  data  from  the  after  period  are  examined. 
Notwithstanding  the  fact  that  the  water  ingestion  of  this  period  was 
reduced  to  the  low  level  of  the  fore  period  the  stools  as  dropped  were  on 
a much  higher  amylolytic  plane  than  were  those  of  the  fore  period.  This 
fact  is  clearly  brought  out  when  it  is  noted  that  the  corresponding  values 
for  the  fore  and  after  periods  were  597  and  1,065  respectively.  We  thus 
see  that  the  stimulation  of  the  factors  which  brought  about  the  increased 
amylolytic  power  of  the  fecal  output  of  the  water  period  was  not  transi- 
tory in  character.  In  other  words  the  influence  of  the  water  was  not 
limited  to  the  time  of  its  ingestion,  but  the  factors  leading  to  an  increased 
fecal  amylolytic  value  were  so  stimulated  by  the  ingestion  of  the  excess 
water  as  to  continue  their  efficiency  even  after  the  water  ration  had  been 
again  reduced  to  that  of  the  fore  period.  Whether  this  pronounced 
increase  in  amylolytic  power  observed  to  accompany  and  follow  the  inges- 
tion of  additional  volumes  of  water  at  meal  time  may  be  properly  inter- 
preted as  indicative  of  an  increased  activity  of  the  pancreatic  function  is 
discussed  in  a later  paragraph. 


7 


In  the  second  experiment  on  the  influence  of  moderate  water  drinking 
(Experiment  2)  E served  as  subject.  All  conditions  were  the  same  as 
those  in  force  in  the  similar  experiment  on  W with  the  exception  that  the 
uniform  diet  was  varied  in  a minor  manner  from  that  ingested  by  W. 
The  data  obtained  from  the  fecal  examinations  of  this  experiment  are 
tabulated  in  Table  2.  In  this  instance  approximate  nitrogen  equilibrium 
was  secured  in  seven  days,  the  nitrogen  balance  showing  a plus  value  of 
0.485  gm.  per  day.  A ten-day  water  period  then  followed  during  which 
500  c.c.  of  water  were  ingested  with  each  meal  in  addition  to  the  uniform 
water  ingestion  of,  the  fore  period.  The  experiment  ended  with  a four- 
day  after  period. 


Table  2. — Fecal  Amylase,  Experiment  2,  Subject  E,  Moderate  Water-Drinking 

FORE  PERIOD SEVEN  DAYS 


Stool 

Feces  - 

Fresh 

Dry 

Amylc 

Total 

•lytic  Value — 1%  Starch 
Per  Gram  Dry 

No. 

(grams) 

(grams) 

(liters) 

Matter  (c.c.) 

1 

88.4 

18.4 

6.3 

343 

2 

30.2 

8.0 

3.9 

491 

3 

179.2 

41.2 

14.0 

341 

4 

193.9 

47.8 

11.3 

237 

5 

76.9 

20.1 

3.7 

184 

6 

207.7 

47.3 

16.3 

345 

7 

121.6 

27.8 

15.3 

548 

8 

44.0 

12.9 

2.2 

171 

Average  per  day. . . 

. . 135.0 

32.0 

10.4 

325 

9 

76.5 

WATER  PERIOD TEN  DAYS 

(1,500  c.c.  H20  daily) 

17.5  7.3 

418 

10 

140.9 

29.4 

31.6 

1,075 

11 

63.8 

18.6 

2.3 

124 

12 

169.0 

41.1 

16.1 

393 

13 

247.5 

47.5 

12.1 

255 

14 

135.3 

31.1 

6.2 

198 

15 

192.4 

37.7 

19.7 

522 

16 

79.2 

19.5 

15.6 

801 

17 

55.9 

15.2 

9.3 

613 

18 

173.5 

43.2 

27.5 

636 

19 

51.5 

14.4 

9.2 

642 

Average  per  day. . . 

..  138.6 

31.5 

15.7 

498 

20 

67.3 

AFTER  PERIOD FOUR 

15.9 

DAYS 

14.0 

878 

21 

117.8 

30.7 

19.9 

649 

22 

147.7 

34.3 

21.8 

637 

23 

145.9 

38.7 

25.8 

667 

24 

31.2 

9.8 

9.4 

961 

Average  per  day. . . 

..  127.5 

32.3 

22.7 

703 

During  the  preliminary  interval  of  low  water 

ingestion  covered  by 

the  fore  period  the  average  daily  total  amylolytic  value  of  the  feces  was 
equivalent  to  10.4  liters  of  1 per  cent,  starch  solution.  Placed  on  a dry- 
matter  basis  we  find  that  the  amylolytic  value  of  the  feces  of  this  period 
may  be  represented  by  325  c.c.  of  1 per  cent,  starch  solution  per  gram  of 
dry  matter.  If  we  now  examine  the  data  for  the  period  during  which  the 
extra  volume  of  water  was  ingested  we  find  that  the  conditions  are 


similar  to  those  already  discussed  in  connection  with  the  experiment  in 
which  W served  as  subject.  In  other  words,  water  caused  an  increase  in 
the  amylolytic  value  of  the  feces.  The  average  total  daily  amylolytic 


% 


8 


value  for  the  water  period  was  15.7  as  against  10.4  for  the  fore  period 
whereas  the  value  on  the  dry  matter  basis  was  498  as  compared  with  a 
value  of  325  for  the  preliminary  interval. 

The  influence  of  the  moderate  water-drinking  in  causing  an  increase 
in  the  amylolytic  power  of  the  feces  was  much  less  pronounced  with  E 
than  with  W.  This  statement  is  borne  out  by  the  observation  that  there 
was  an  increase  of  nearly  100  per  cent,  in  the  case  of  W as  against  an 
increase  of  little  more  than  50  per  cent,  in  the  case  of  E.  However,  when 
we  examine  the  data  for  the  after  periods  of  the  two  experiments  in 
question,  we  note  that  the  amylolytic  value  in  E?s  experiment  was  above 
that  of  the  water  period,  whereas  in  W’s  experiment  this  value  was 
slightly  lower  than  that  of  the  water  period.  By  calculation  we  see  that 
the  amylolytic  value  for  the  after  period  of  the  test  on  W was  increased 
only  80  per  cent,  above  that  of  the  fore  period  whereas  the  increase  in  the 
test  on  E was  about  120  per  cent.  It  would  seem,  therefore,  that  the 
water  caused  a more  pronounced  immediate  stimulation  in  the  case  of 
W than  in  that  of  E but  that  the  influence  of  the  stimulation  was  more 
persistent  in  the  case  of  E than  in  that  of  W. 


Table 

3.- 

-Fecal 

Amylase, 

Experiment  3, 

Subject  E,  Copious 

Water-Drinking 

FORE  PERIOD — 

-SIX  DAYS 

— Feces  - 

Amylolytic 

Value — 1%  Starch 

Stool 

Fresh 

Dry 

Total 

Per  Gram  Dry 

No. 

(grams) 

(grams) 

(liters) 

Matter  (c.c.) 

1 

35.2 

10.2 

3.8 

375 

66.0 

18.8 

8.6 

458 

3 

202.2 

49.8 

23.6 

473 

4 

3 29.2 

32.2 

14.9 

464 

5 

161.3 

34.6 

8.8 

256 

6 

171.8 

38.8 

15.8 

406 

7 

34.6 

9 4 

4.2 

448 

Average 

per 

day. . . 

. . 133.4 

32.3 

13.3 

412 

WATER  PERIOD— 

-FIVE  DAYS 

(4,000  c.c.  H20  daily) 

8 

90.3 

18.2 

10.5 

578 

9 

37.2 

10.2 

8.3 

816 

10 

249.4 

58.8 

26.5 

451 

11 

74.7 

3 9.9 

6.1 

306 

12 

258.0 

38.2 

342.3 

8,961 

13 

52.6 

15.9 

7.8 

491 

Average 

per 

day. . . . 

. . 152.5 

32.2 

80.3 

2,494 

AFTER  PERIOD THREE  DAYS 

14 

128.3 

29.6 

12.3 

414 

15 

86.4 

21.0 

15.6 

743 

16 

206.5 

44.5 

25.0 

561 

17 

50.6 

5.6 

7.0 

1,257 

Average 

per 

day 

. 157.3 

33.6 

20.0 

595 

2.  Copious  Water-Drinking. — The  amylolytic  power  of  the  feces  was 
studied  in  but  one  experiment  in  which  copious  water  drinking  was  prac- 
ticed at  meal  time.  In  this  experiment  (3)  E served  as  subject.  All  the 
experimental  conditions  were  similar  to  those  in  force  with  this  same 
subject  in  the  second  experiment  on  moderate  water  drinking.  In  a fore 
period  covering  an  interval  of  six  days  a very  satisfactory  balance  for 
income  and  outgo  of  nitrogen  was  obtained,  the  data  indicating  a plus 
balance  of.  only  0.03  gm.  The  data  from  this  experiment  are  given  in 
Table  3.  An  examination  of  that  table  will  show,  in  the  first  place,  that 


9 


the  average  total  daily  amylolytic  value  for  the  fore  period  was  13.3, 
whereas  the  amylolytic  value  on  a dry  matter  basis  was  412.  These  values 
simply  mean,  as  heretofore  explained,  that  the  average  output  of  feces 
per  day  during  the  six-day  fore  period  possessed  the  power  to  hydrolyze 
13.3  liters  of  1 per  cent,  starch  solution,  whereas  1 gm.  of  dry  feces 
possessed  the  power  of  transforming  412  c.c.  of  such  a starch  solution. 

The  water  period  of  this  experiment  was  five  days  in  duration  as 
against  the  ten-day  periods  utilized  in  the  experiments  already  discussed. 
However,  the  daily  ingestion  of  water  was  much  higher  in  this  instance 
than  in  either  of  the  afore  mentioned  studies.  In  the  moderate  water- 
drinking  tests  we  caused  the  subjects  to  ingest  500  c.c.  additional  at  each 
meal  or  a total  daily  ingestion  of  1,500  c.c.  in  excess  of  that  customarily 
taken.  In  certain  other  water-drinking  studies  made  by  us  (Fowler  and 
Hawk1;  Mattill  and  Hawk22;  also  Wills  and  Hawk,  unpublished),  in 
which  the  influence  of  copious  amounts  of  water  was  under  investigation, 
the  total  volume  of  water  added  daily  to  the  normal  ration  had  been 
3,000  c.c.  However,  in  the  present  investigation  we  were  dealing  with  a 
subject,  E,  who  was  accustomed  to  drinking  rather  larger  volumes  of 
water  than  individuals  ordinarily  ingest  and  for  this  reason  his  water 
ration  was  fixed  at  4,000  c.c.  per  day  during  the  water  period  in  an 
attempt  to  place  the  daily  volume  at  such  a figure  as  should  be  copious 
for  his  organism.  He  felt  no  personal  discomfort  at  any  time  during  the 
period  in  which  this  excessive  amount  of  water  was  being  daily  introduced 
into  his  system. 

Under  the  influence  of  copious  water-drinking  through  a five-day 
period,  the  amylolytic  activity  of  E?s  feces  was  increased  in  an  extremely 
emphatic  manner.  Particularly  was  this  true  of  Stool  12,  passed  on  the 
fourth  day  of  the  period.  Expressing  the  amylolytic  values  in  the  same 
manner  as  heretofore  followed  we  find  that  the  average  amylolytic  power 
of  the  feces  was  equivalent  to  a force  sufficient  to  hydrolyze  80.3  liters  of 
1 per  cent,  starch  solution.  Furthermore,  if  we  transfer  the  comparison 
to  the  dry-matter  basis  we  learn  that  1 gram  of  dry  matter  represents 
sufficient  amylolytic  activity  to  hydrolyze  2,494  c.c.  of  1 per  cent,  starch 
solution.  When  we  compare  these  volumes  with  those  obtained  during 
the  fore  period  we  are  astounded  at  the  very  material  increase  in  the 
amylolytic  activity  which  has  taken  place  under  the  influence  of  the 
copious  water-drinking.  For  example,  if  we  consider  the  dry-matter  basis 
we  see  that  the  amylolytic  value  has  increased  from  412  to  2,494,  a most 
surprising  increase  of  more  than  600  per  cent,  in  the  amylolytic  activity. 
The  question  as  to  the  proper  interpretation  of  this  finding,  in  so  far  as 
it  relates  to  the  functional  activity  of  the  pancreas,  is  discussed  later  on. 
In  this  experiment  for  the  first  time,  we  observed  an  amylolytic  value  for 
the  after  period  which  was  lower  than  that  of  the  water  period.  Evi- 


10 


dently  the  stimulation  of  the  copious  water-drinking  was  so  pronounced 
in  character  in  this  particular  instance  as  to  rather  militate  against  any 
further  added  stimulation  after  the  organism  was  released  from  the  imme- 
diate influence  of  the  water  ingestion.  That  the  influence  of  the  water 
extended  somewhat  beyond  the  time  interval  during  which  it  was  actually 
being  ingested  is  gathered  from  the  recorded  average  amylolytic  values 
for  the  after  period.  These  values  are  nearly  45  per  cent,  above  those  of 
the  fore  period  thus  indicating  an  after  effect  which  persisted  for  a time, 
at  least,  after  the  period  of  high  water  intake  had  closed. 

INTERPRETATION  OF  FINDINGS 

The  foregoing  discussion  of  the  various  experiments  embraced  in  this 
study  has  indicated  clearly  that  when  either  small  or  large  volumes  of 
water  above  those  customarily  ingested  were  taken  at  meal  time  that  the 
resultant  fecal  matter  possessed  greater  amylolytic  activity  than  did  the 
feces  passed  during  periods  in  which  these  extra  volumes  of  water  were 
not  taken.  How  is  this  finding  to  be  interpreted?  It  has  been  estab- 
lished that  an  increase  in  the  activity  of  the  pancreatic  function  will  be 
indicated  by  the  appearance  of  an  added  excretion  of  pancreatic  amylase 
in  the  feces.  This  fact  would  of  itself  cause  the  feces  in  question  to 
possess  an  increased  power  to  hydrolyse  starch  solution.  On  such  a basis 
Wohlgemuth  has  suggested  a method  for  the  quantitative  determination 
of  the  amylolytic  power  of  the  feces.  This  power,  according  to  Wohlge- 
muth, is  due  entirely  to  the  ability  of  dilute  solutions  of  the  fecal  amylase 
to  transform  starch  into  substances  which  no  longer  give  a blue  color 
with  iodin.  From  this  point  of  view,  therefore,  we  may  interpret  our 
results  as  indicating  that  the  pancreatic  function  has  been  greatly  stimu- 
lated under  the  influence  of  the  water-drinking  at  meal-time  and  that 
consequently  larger  quantities  of  pancreatic  amylase  are  present  in  the 
feces  passed  during  the  water  period,  thus  giving  to  the  fecal  matter  a 
higher  amylolytic  power  than  that  possessed  by  stools  dropped  during 
previous  periods  in  which  the  water  ingestion  was  minimal. 

We  began  our  investigation  under  the  impression  that  Wohlgemuth’s 
method  would  furnish  us  with  accurate  data,  and  nothing  occurred  in  the 
course  of  the  experiments  to  cast  any  doubt  on  the  validity  of  Wohlge- 
muth’s claim  until  an  examination  was  made  of  the  final  stool  (17)  in 
the  water  period  of  Experiment  1.  Here  for  the  first  time  in  the  course 
of  our  studies  the  entire  series  of  seven  tubes  of  1 per  cent,  starch  solution 
showed  complete  digestion.  As  soon  as  this  fact  was  determined  another 
series  of  ten  tubes  was  prepared  and  again  the  entire  series  was  com- 
pletely digested.  Hot  caring  to  rely  on  data  obtained  from  further 
extracts  because  of  the  possibility  of  changes  having  taken  place  in  the 
feces,  we  examined  no  further  series.  A similar  observation  was  made  in 


11 


connection  with  Stool  12  in  Experiment  3.  In  onr  search  as  to  the  cause 
of  this  surprisingly  pronounced  increase  in  the  amylolytic  activity  of  the 
stools  mentioned,  we  observed  that  they  each  possessed  a very  pronounced 
acid  reaction.  We  were  familiar  with  the  findings  of  Chittenden  and 
Griswold,23  which  were  later  confirmed  by  Vernon24  to  the  effect  that 
amylase  (salivary)  is  inactive  in  the  presence  of  0.009  per  cent,  hydro- 
chloric acid.  At  the  same  time  we  were  familiar  with  the  further  fact 
that  it  requires  an  acid  concentration  much  above  0.009  per  cent,  hydro- 
chloric acid  to  hydrolyze  1 per  cent,  starch  solutions  under  the  conditions 
of  our  experiment.  Furthermore,  in  the  experiments  of  Vernon  to  which 
reference  has  already  been  made,  as  well  as  in  others  by  Schierbeck,23  it 
has  been  demonstrated  that  certain  proper  acid  concentrations  below  0.009 
per  cent,  hydrochloric  acid  will  facilitate  the  activity  of  amylase.  This 
power  was  shown  to  be  possessed  by  organic  acids  as  well  as  by  inorganic 
acids.  In  the  case  of  hydrochloric  acid  a concentration  of  0.004  per  cent, 
was  found  to  cause  an  increase  of  400  per  cent,  in  the  amylolytic  power 
of  the  enzyme,  whereas  0.0083  per  cent,  lactic  acid  increased  the  amylo- 
lysis  nearly  500  per  cent. 

In  light  of  the  above  findings,  we  might  interpret  the  fivefold  increase 
in  the  amylolytic  power  of  acid  Stool  17  of  Experiment  1 and  the  sixfold 
increase  in  the  amylolytic  power  of  acid  Stool  12  of  Experiment  3 as  due 
to  the  stimulating  influence  of  the  acid  reaction  of  the  sodium  chi  or  id 
extracts  of  the  feces  on  the  contained  amylase.  It  obviously  needs  no 
argument  to  support  the  claim  that  the  acidity  of  these  extracts  could 
not  of  itself  have  caused  the  hydrolysis  of  the  1 per  cent,  starch  solutions. 
Even  had  the  feces  possessed  a surprisingly  high  acid  concentration,  this 
concentration  must  of  necessity  have  been  so  much  lowered  by  the  time 
the  series  of  dilutions  of  the  salt  solution  extracts  had  been  made  as  to 
render  the  acidities,  of  the  latter  part  of  the  series,  at  least,  of  no  influence 
in  so  far  as  their  power  to  hydrolyze  starch  is  concerned.  It  is  entirely 
possible  that  the  amylase  was  passed  into  the  intestine  in  increased 
amount  on  the  days  in  question  under  the  influence  of  the  high  water 
intake,  that  it  there  fulfilled  its  function  of  bringing  about  an  augmented 
anrylolysis  and  that  subsequently  the  reaction  of  the  surrounding  media 
was  so  altered  by  certain  factors  as  to  render  impossible  any  further 
activity  of  the  enzyme.  Therefore  the  reaction  in  which  the  enzyme  was 
finally  excreted  from  the  organism  by  way  of  the  feces  was  not,  of  neces- 
sity,  the  reaction  which  the  intestinal  substrate  of  the  pancreatic  amylase 
possessed.  Be  that  as  it  may,  however,  it  is  of  course  impossible  to  come 
to  any  definite  conclusion  as  to  the  extent  of  the  stimulation  of  the 
activity  of  the  pancreatic  function  on  the  basis  of  data  collected  from  acid 

23.  Chittenden  and  Griswold:  Am.  Chem.  Jour.,  1881,  iii,  305. 

24.  Vernon:  Jour.  Physiol.,  1902,  xxvii,  174. 

25.  Schierbeck:  Skand.  Arch.  f.  Physiol.,  1892,  iii,  344. 


12 


stools  by  means  of  the  Wohlgemuth  method.  This  being  so,  we  have 
suggested  a modification  of  that  method  which  should  obviate  the  trouble- 
some features  associated  with  the  examination  of  stools  which  possess  a 
pronounced  acid  or  alkaline  reaction.  This  method  appears  elsewhere.20 

If  we  examine  the  data  in  Tables  1,  2 and  3 it  will  be  seen  that  the 
complete  elimination  of  all  consideration  of  the  acid  stools  in  question 
which  gave  evidence  of  the  surprisingly  pronounced  amylolytic  power,  will 
not  prevent  us  from  drawing  the  conclusion  that  the  activity  of  the  pan- 
creatic function  was  stimulated  under  the  influence  of  water-drinking  at 
meal  time.  Certain  other  experiments  may  be  cited  as  tending  to  sub- 
stantiate the  theory  of  an  increased  activity  of  the  pancreatic  function 
under  the  influence  of  water-drinking  with  meals.  For  example,  Pawlow26 
has  shown  that  if  150  c.c.  of  water  be  introduced  into  the  stomach  of  a 
dog  possessing  a pancreatic  fistula,  that  the  flow  of  juice  begins  after  an 
interval  of  two  or  three  minutes  and  in  case  there  is  already  present  a 
distinct  flow  of  juice  that  the  output  is  distinctly  increased.  That  this 
flow  of  juice  was  not  brought  about  through  the  stimulation  of  acid  chyme 
entering  the  duodenum  was  shown  from  the  fact  that,  if  the  stomach  of 
the  animal  be  emptied,  the  contents  of  the  organ  are  found  to  be  neutral 
or  faintly  alkaline  in  reaction.  This  experiment  by  Pawlow  indicates 
clearly,  then,  that  water  introduced  into  the  stomach  causes  a direct 
stimulation  of  the  nervous  mechanism  of  the  pancreas  which  is  followed 
by  an  outpouring  of  pancreatic  juice.  As  further  basis  for  our  belief  that 
water  ingestion  stimulates  the  pancreas,  we  would  cite  certain  other 
experiments  of  Pawlow27  and  the  earlier  work  of  Heidenhain28  and  Ssa- 
nozki.28  The  more  recent  experiments  of  Foster  and  Lambert29  may  also 
be  adduced  as  furthering  this  claim.  These  various  investigators  just 
enumerated  have  shown  that  an  increased  flow  of  gastric  juice  follows  the 
entrance  of  water  into  the  stomach.  The  studies  of  Foster  and  Lambert 
go  even  farther  than  this  and  show  most  clearly  that  there  is  not  only 
an  increased  flow  of  juice  but  that  this  increased  volume  possesses  a 
higher  acid  concentration  than  that  possessed  by  juice  secreted  under 
other  conditions.  We  have  obtained  results  in  this  laboratory30  which  go 
to  verify  the  findings  of  Foster  and  Lambert.  On  the  basis  of  the  well- 
established  theory  as  to  the  mechanism  of  pancreatic  secretion  as  evolved 
by  Bayliss  and  Starling31  we  may  logically  expect  a pronounced  increase 
in  the  flow  of  pancreatic  juice  to  follow  the  entrance  of  the  strongly  acid 
chyme  into  the  duodenum.  That  the  pancreas  is  more  active  under  a 

26.  Pawlow:  The  Work  of  the  Digestive  Glands,  Second  Edition,  1910,  p.  144. 

27.  Pawlow:  The  Work  of  the  Digestive  Glands,  p.  112. 

28.  Quoted  by  Pawlow,  p.  112. 

29.  Foster  and  Lambert:  Jour.  Exper.  Med.,  1908,  x,  820. 

30.  Wills  and  Hawk:  Proc.  Soc.  of  Biol.  Chem.,  1911. 

31.  Bayliss  and  Starling:  Jour.  Physiol.,  1902,  28. 


13 


high  water  ingestion  is  also  evidenced  from  other  experiments  made  in 
this  laboratory22  which  have  demonstrated  a more  complete  digestion  and 
absorption  of  ingested  fats  and  carbohydrates  under  the  influence  of 
water-drinking  at  meal  time. 

It  has  further  been  found  by  us32  that  the  carbohydrate  content  of 
the  actual  samples  of  feces  listed  in  Tables  1,  2 and  3 of  this  article  was 
lower  during  the  water  periods  than  during  the  period  of  minimal  water 
ingestion.  This  fact  demonstrated  definitely  then  that  the  intestinal 
content  of  these  subjects  was  of  a higher  amylase  concentration  during 
the  period  of  increased  water  intake.  Other  things  being  equal  we  would 
expect  a fair  degree  of  uniformity  between  the  availability  of  ingested 
carbohydrate  and  the  content  of  fecal  amylase.  The  observation  of  Brad- 
ley33 to  the  effect  that  pancreatic  lipase  possesses  greater  fat-splitting 
power  when  its  reaction  mixtures  are  diluted  with  several  volumes  of 
water  also  goes  far  toward  a substantiation  of  the  theory  of  a pancreas 
activated  through  water  ingestion. 

We  have  much  to  learn  as  to  the  alterations  in  the  reaction  of  the 
intestine  under  different  conditions.  In  the  case  in  point  we  have  an 
excessive  quantity  of  hydrochloric  acid  passed  into  the  intestine,  under 
the  influence  of  the  high  water  intake,  a quantity  which  probably,  at 
times,  is  far  in  excess  of  that  which  may  be  neutralized  by  the  pancreatic 
juice  and  bile.  The  greater  portion  of  this  excess  acid  we  may  suppose 
to  be  neutralized  by  the  ammonia  which  is  produced  in  the  deamidation 
of  protein  material.  This  acid  would  therefore  appear  in  the  urine  as 
ammonium  chlorid.  However,  if  there  should  occur  a very  copious  out- 
pouring of  hydrochloric  acid  at  a time  when  there  was  but  little  protein 
material  available  for  deamidation  and  consequently  a minimal  amount  of 
ammonia  present  for  the  neutralization  of  such  excess  acid,  it  is  entirely 
possible  that  we  might  have  an  acid  reaction  due  to  hydrochloric  acid 
extending  the  entire  length  of  the  large  intestine.  In  that  event  the  stools 
would  probably  possess  a relatively  high  acid  concentration.  If  the  food 
residues  remained  in  the  intestine  for  an  interval  shorter  than  the  normal 
because  of  increased  peristalsis,  diarrhea,  etc.,  acid  stools  might  also 
result. 

The  normal  reaction  of  the  intestinal  contents  is  acid  down  to  the 
ileocecal  valve  under  normal  conditions.  Furthermore,  carbohydrates 
which  escape  absorption  may  undergo  acid  fermentation  through  the 
action  of  certain  acid-forming  bacteria  such,  for  example,  as  Bacillus  coli 
and  Bacterium  welchii.  A number  of  organic  acids  are  formed  which 
include  acetic  acid,  lactic  acid,  butyric  acid  and  succinic  acid.  The  extent 
to  which  this  acid  reaction  may  be  carried  is  indicated  by  Macfadyen, 


32.  Mattill  and  Hawk:  Unpublished  results. 

33.  Bradley:  Journal  Biol.  Chem.,  1910,  viii,  251. 


14 


Nencki  and  Sieber’s34  finding  of  an  acidity  at  the  ileocecal  valve  equiva- 
lent to  0.1  per  cent,  acetic  acid.  This  observation  was  made  on  a patient 
with  a fistula  at  the  end  of  the  small  intestine. 

SUMMARY 

The  problem  studied  was  the  activity  of  the  pancreatic  function  under 
the  influence  of  water-drinking  at  meal  time.  Normal  men  were  used 
as  subjects  and  were  required  to  ingest  a diet  uniform  in  all  respects 
from  day  to  day.  Three  experiments  were  made,  each  experiment  being 
divided  into  three  periods;  a fore  period  in  which  the  subject  was  brought 
into  nitrogen  equilibrium  through  the  ingestion  of  the  uniform  diet  sup- 
plemented by  a minimal  water  ingestion;  a water  period  during  which  the 
water  ingestion  at  meal  time  was  increased  from  1,500  to  4,000  c.c.  per 
day ; and  an  after  period  in  which  the  dietary  conditions  of  the  fore  period 
prevailed.  Two  of  the  experiments  were  on  the  influence  of  moderate 
water-drinking  and  in  these  the  volume  of  the  extra  water  ingested  during 
the  water  period  was  1,500  c.c.  per  day  or  500  c.c.  per  meal.  In  the  third 
experiment  the  influence  of  copious  water-drinking  was  studied  and  in 
this  instance  the  subject  was  required  to  ingest  on  each  day  of  the  water 
period  4,000  c.c.  above  that  ingested  in  the  fore  and  after  periods. 

The  amylolytic  activity  of  the  feces,  denoting,  according  to  Wohl- 
gemuth, the  content  of  pancreatic  amylase  present  in  the  feces,  was  taken 
as  the  index  of  the  activity  of  the  pancreatic  function.  The  amylolytic 
values  for  the  stools  dropped  during  the  periods  of  moderate  and  copious 
water-drinking  at  meal-time  were  much  higher  than  the  similar  values  as 
determined  for  the  stools  dropped  during  the  periods  of  minimal  water 
ingestion.  This  finding  may  be  interpreted  as  indicating  that  the  drink- 
ing of  water  with  meals  had  stimulated  the  pancreas.  We  make  this 
interpretation  with  certain  reservations  regarding  conditions  not  con- 
trolled by  the  Wohlgemuth  method.  Two  stools  were  encountered  in  the 
course  of  the  investigation  which  exhibited  a pronounced  acid  reaction. 
Inasmuch  as  the  basic  principle  of  Wohlgemuth’s  method  is  the  hydrolysis 
of  starch  solutions  through  the  medium  of  unneutralized  fecal  extracts, 
the  method  does  not  give  dependable  data  as  to  the  amylase  content  when 
such  stools  are  under  examination.  This  is  true  especially  in  view  of  the 
fact  that  it  has  been  shown  that  0.004  per  cent,  hydrochloric  acid  will 
increase  the  activity  of  amylase  400  per  cent.,  whereas  0.009  per  cent, 
hydrochloric  acid  will  cause  absolute  inhibition.  The  power  of  feces 
extracts,  therefore,  to  hydrolyze  starch  cannot  be  taken  as  a measure  of 
the  amylase  present  unless  precautions  are  taken  to  neutralize  the  fecal 
extracts  and  then  make  the  conditions  uniform  for  the  action  of  the 
enzyme  if  present. 

34.  Macfadyen,  Nencki  and  Sieber:  Arch.  f.  exper.  Patliol.  u.  Pharmacol., 
1891.  xxviii,  311. 


15 


On  the  basis  of  the  data  gathered  in  this  and  in  associated  investiga- 
tions made  in  our  laboratory  and  elsewhere,  we  are  prepared  to  draw  the 
general  conclusion  that  the  ingestion  of  quantities  of  water  at  meal-time 
ranging  in  volume  from  y2  to  1%  liter  stimulate  the  pancreatic  function 
in  two  ways : first,  a direct  stimulation  of  the  nervous  mechanism  of  the 
pancreas  brought  about  while  the  water  is  still  in  the  stomach  and,  second, 
an  indirect  stimulation  brought  about  on  the  entrance  of  the  increased 
volume  of  acid  chyme  into  the  duodenum.  The  drinking  of  water  with 
meals  ought  therefore  to  bring  about  a more  rapid  and  complete  digestion 
and  absorption  of  the  fat  and  carbohydrate  constituents  of  the  diet,  two 
observations  verified  by  experimentation  in  our  laboratory. 


